1 research outputs found
Reconstruction of a human cornea by the self-assembly approach of tissue engineering using the three native cell types
Purpose: The purpose of this study was to produce and characterize human tissue-engineered corneas reconstructed using
all three corneal cell types (epithelial, stromal, and endothelial cells) by the self-assembly approach.
Methods: Fibroblasts cultured in medium containing serum and ascorbic acid secreted their own extracellular matrix and
formed sheets that were superposed to reconstruct a stromal tissue. Endothelial and epithelial cells were seeded on each
side of the reconstructed stroma. After culturing at the air-liquid interface, the engineered corneas were fixed for histology
and transmission electron microscopy (TEM). Immunofluorescence labeling of epithelial keratins, basement membrane
components, Na+/K+-ATPase α1, and collagen type I was also performed.
Results: Epithelial and endothelial cells adhered to the reconstructed stroma. After 10 days at the air-liquid interface, the
corneal epithelial cells stratified (4 to 5 cell layers) and differentiated into well defined basal and wing cells that also
expressed Na+/K+-ATPase α1 protein, keratin 3/12, and basic keratins. Basal epithelial cells from the reconstructed
epithelium formed many hemidesmosomes and secreted a well defined basement membrane rich in laminin V and collagen
VII. Endothelial cells formed a monolayer of tightly-packed cells and also expressed the function related protein Na+/K
+-ATPase α1.
Conclusions: This study demonstrates the feasibility of producing a complete tissue-engineered human cornea, similar
to native corneas, using untransformed fibroblasts, epithelial and endothelial cells, without the need for exogenous
biomaterial